Human papillomaviruses (HPVs) are associated with a number of human malignancies including cervical cancer, other anogenital cancers, and a subset of head and neck cancers. The goals of this research project are to continue to further define and understand the viral functions and virus:host interactions that mediate HPV-associated disease, from the point of infection to development of HPV-associated malignancy. Over the current funding period, the laboratories of Drs. Lambert and Ahlquist have made a number of important contributions to our understanding of HPV-associated disease, with the development of novel mouse models for HPV-associated. malignancy, gene expression profiling of HPV-associated human cancers, identification of useful biomarkers for HPV-associated cancers, development of a highly efficient means for producing HPV virion particles and its use in studies evaluating host interactions in early stages of HPV-infection, and studies on the individual roles of HPV genes in the viral life cycle. In this new project, Drs. Lambert and Ahlquist together will characterize the nature and mechanisms of virus:host interactions in the context of HPV infections and HPV-associated malignancy. The specific aims are: (1) to investigate viral:host interactions in the early steps of viral infection (i.e., binding, entry and establishment); (2) to define for mechanistic and epidemiological analysis the changes in human gene and microRNA expression that characterize each stage in the long progression from HPV infection to cervical cancer; and (3) to further characterize mouse models for HPV-associated cervical cancer and to use these mouse models to investigate the role of particular cellular genes, including those identified in Aim 2, in cervical cancer. As in all other projects of this program-project grant, the results should have significant implications for diagnosis, prognosis, and/or therapy of persistent tumor virus infection and virus-associated cancers.